In Vitro Antioxidant Activity of Ethanolic and Chloroform Extracts of Tagetes erecta Flowers
Nirmala Shinde1*, R.A. Saindane1, S.K.
Bhosale1 and Shreenivas Deshpande2
1Department of Pharmaceutical
Chemistry, S. M. B. T. College of Pharmacy, Dhamangaon,
Igatpuri, Nashik-422403 (M. S.)
2Department of Pharmaceutical
Chemistry, H. S. K. College of Pharmacy, Bagalkot,
(Karnataka)
ABSTRACT:
To evaluate the in-vitro
free radical scavenging activity of flowers of Tagetes erecta, ethanolic and
chloroform extracts of flowers of Tagetes erecta were prepared with successive extraction in soxhlet apparatus. Each extract was selected to study the antioxidant
activity by iron Chelation method and hydrogen
peroxide scavenging method. The phytochemical studies
were also carried out for both the extracts to evaluate the presence of different
phytochemical constituents. The percentage of iron
Chelating activity for Ethanolic and Chloroform
Extract were found to be 53.71±14.51% and 55.77±9.67% respectively. The
percentage of hydrogen peroxide scavenging for Ethanolic and Chloroform extracts of were found 9.51±2.49%
and 13.03±7% respectively. The comparison was done with the standard Ascorbic
acid. The present study revealed that both the
extracts effectively show their ability to scavenge the free radicals as well
as to bind metal ions needed for catalysis of reactive oxygen
generation.
KEYWORDS: Tagetes erecta, antioxidant activity, Iron Chelating Activity,
Scavenging of Hydrogen Peroxide.
INTRODUCTION:
Antioxidants are substances that are capable of
counteracting the damaging effects of the physiological process of oxidation in
animal tissues. Antioxidants may be nutrients (vitamins and minerals) or
enzymes. Oxidative stress occurs when the production of harmful molecules
called free radicals is beyond the protective capability of the antioxidant
defenses. Free radicals are chemically active atoms
or molecular fragments that have a charge due to an excess or deficient number
of electrons. Examples of free radicals are the superoxide anion, hydroxyl
radical, transition metals such as iron and copper, nitric acid, and ozone.
Free radicals containing oxygen, known as reactive oxygen species (
Tagetes erecta (Compositae), also
called American marigolds, are tall stout plants that grow to 0.9 m in height.6,
7. It is commonly used in the treatment of fungal infections, in the
preparation of some herbaceous and floral perfumes and to deter house flies.
Essential oil of leaves completely found to inhibit the growth of Fusarium oxysporum and
Trichophyton mentagrophytes
and the mycelial growth of Pythium
aphanidermatatum. When tested against 40 strains
of bacteria and fungi, essential oil of Tagetes erecta was found to have a 100% inhibitory effect against
Gram-positive bacteria and a 95% inhibitory effect against fungi.
By
keeping in mind the so many properties exhibited by Tagetes erecta,
in the
present work an attempt is made to study antioxidant activity of chloroform and
ethanolic extract of the flowers of Tagets erecta.
MATERIALS AND METHODS:
Collection of Plant:
The
flowers of Tagetes erecta was
collected from the farms in area nearby Nashik. The
collected plant material was authentified from the
botanist and shade dried.
Extraction of Plant material:
The
shade dried flowers of Tagetes erecta were
powdered and then extracted with the ethanol and chloroform by using Soxhlet apparatus.
Phytochemical
Evaluation of extract8, 9.
Both
the extracts were evaluated for the presence of different phytochemical
constituents.
The
results are as shown in the Table No. 1.
Table No.1: Presence of different Phytochemical constituents in ethanolic and chloroform extracts
|
Sr. No. |
Phytochemical Constituent |
Presence/Absence of Phytochemical
Constituent |
|
|
Ethanolic extract |
Chloroform extract |
||
|
1 |
Alkaloids |
+ |
+ |
|
2 |
Carbohydrates |
+ |
+ |
|
3 |
Cardiac Glycosides |
_ |
+ |
|
4 |
Anthraquinones |
+ |
+ |
|
5 |
Flavanoids |
+ |
+ |
|
6 |
Saponins |
+ |
_ |
+
indicates presence of phytochemical constituent.
-
indicates absence of phytochemical constituent.
Screening of antioxidant activity:
Both
the extracts were screened for their anti-oxidant activity by following methods
A) Iron Chelating Activity:
The
reaction mixture containing 1 ml 0.05% o-phenathroline
in methanol, 2 ml ferric chloride 200 mM and 2 ml
various concentrations of the test compound was incubated at ambient
temperature for 10 min and the absorbance of the same was measured at 510 nm.
The experiment was performed in duplicate.10
absorbance (control) – absorbance (sample)
%Chelation =
-------------------------------------------------- X 100
absorbance (control)
B) Scavenging of hydrogen peroxide:
A
solution of hydrogen peroxide (40 mMol L–1)
was prepared in phosphate buffer
(pH 7.4). Different concentrations (250–2500 mg mL–1) of
extract were added to the hydrogen peroxide solution (40 mmol
L–1, 0.6 mL). Absorbance of hydrogen
peroxide at 230 nm was determined after 10 min against a blank solution containing
phosphate buffer without hydrogen peroxide. Percentage scavenging of hydrogen
peroxide of the extracts and standard compounds was calculated.11
absorbance (control) – absorbance
(sample)
%Scavenging = -------------------------------------------- X 100
Absorbance (control)
RESULTS:
In
the present investigation, the antioxidant activity of ethanolic
and chloroform Extracts of Tagetes erecta flowers was studied in two in vitro antioxidant
models.
Results
of iron chelating activity are shown in table no.2.
Results
for hydrogen peroxide scavenging activity are shown in table no.3.
Table No. 2: % Chelation
by Ethanolic extract, Chloroform extract and Ascorbic
acid solution for Iron Chelation Activity.
|
Test compound |
% Chelation |
||||
|
Concentration(mg/ml) |
50 |
100 |
150 |
200 |
250 |
|
Ethanolic extract |
17.86 |
52.08 |
41.10 |
30.48 |
19.6 |
|
Chloroform extract |
43.32 |
29.85 |
38.30 |
37.38 |
18.45 |
|
Ascorbic acid solution |
77.35 |
58.69 |
48.56 |
32.87 |
39.00 |
Table No. 3: % scavenging of hydrogen
peroxide by Ethanolic extract, Chloroform extract and
Ascorbic acid solution.
|
Test compound |
% inhibition |
||||||
|
Concentration(mg/ml) |
50 |
100 |
150 |
200 |
250 |
300 |
350 |
|
Ethanolic extract |
8.89 |
12.93 |
10.29 |
6.97 |
7.19 |
12.59 |
7.69 |
|
Chloroform
extract |
17.5 |
13.45 |
11.02 |
4.49 |
2.89 |
22.61 |
19.22 |
|
Ascorbic
acid solution |
39.2 |
57.41 |
53.33 |
35.48 |
58.31 |
63.08 |
75.34 |
DISCUSSION:
In
both the models tested, the antioxidant activity of ethanolic
and chloroform extracts of Tagetes erecta flowers was studied in relation to that of
ascorbic acid which is a known standard
potent antioxidant. Even though found to be less potent than Ascorbic Acid, our
study has reported the ability of Ethanolic and
Chloroform Extracts of Tagetes erecta
flowers to act as an antioxidant by several mechanisms viz., scavenging of
reactive oxygen species or their precursors, binding metal ions needed for
catalysis of reactive oxygen generation and by the up-regulation of endogenous
antioxidant defenses.
The
results of this study therefore justify the therapeutic potential of ethanolic and chloroform extracts of Tagetes erecta flowers in traditional system of
medicine, especially in diseases where the antioxidant property is an important
component. The structure activity relationship, synergistic mode of action and
its relative importance in different mechanisms will provide deeper insights in
finding out better and safer therapeutics.
ACKNOWLEDGEMENT:
The authors
are thankful to S. M. B. T. College of
Pharmacy, Dhamangaon, for providing all the necessary
facilities required to carry out the work.
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Received
on 09.01.2010
Accepted on 20.03.2010
© A&V Publication all right reserved
Research Journal of Pharmacognosy and Phytochemistry.
2(4): July-Aug. 2010, 324-326